Vector-Borne Diseases: Biology Of Vector Host Relationship

媒介传播疾病：媒介宿主关系的生物学

基本信息

批准号：
10272070
负责人：
Jose Ribeiro
金额：
$ 110.63万
依托单位：
NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10272070
关键词：
Adult Aedes African Trypanosomiasis Amino Acid Sequence Anti-Inflammatory Agents Area Arthropods Binding Biogenic Amines Bioinformatics Biological Biological Assay Biology Blood Blood Proteins Blood coagulation Brazil CRISPR/Cas technology Cell Line Chemicals Chicago Coagulation Process Code Collaborations Complement Complex Crithidia fasciculata Culicidae Databases Dictyoptera Disease Embryonic Development Eukaryotic Cell Evolution Family Fat Body Female Future Gene Expression Profiling Gene Family Genes Genome Goals Greek Hemorrhage Hemostatic Agents Hemostatic function Human Immune response Impairment Individual Injury Innate Immune Response Insecta Investigation Juvenile Hormones Knock-out Laboratories Lead Learning Leishmaniasis Leukotrienes Magic Mammals Maps Mass Spectrum Analysis Medical Methodology Methods Molecular Molecular Biology Names National Institute of Allergy and Infectious Disease Organism Ovoviviparity Paper Parasites Pathology Patients Peptides Pharmacologic Substance Pharmacology Physiological Physiological Processes Plasmodium falciparum Platelet aggregation Process Property Protein Family Proteins Proteome Protozoan Genome Publications Recombinants Research Research Personnel Rhipicephalus sanguineus Rickettsia Infections Role Saliva Salivary Salivary Glands Salivary Proteins Sand Flies Site System Techniques Tick-Borne Encephalitis Virus Ticks Time Transcript Transcriptional Regulation Tsetse Flies Universities Vaccines Vasodilator Agents Vector-transmitted infectious disease Vertebrates Virginia Virus Work arm cDNA Library design feeding fly human disease immunoreaction immunoregulation in vitro testing in vivo insight interest male member milk production novel pathogen prevent programs protein expression skills success sucking tick feeding tick saliva transcriptome transmission process vaccine development vasoconstriction vector viral transmission

项目摘要

Because host hemostasis (the physiological process that prevents blood loss, consisting of platelet aggregation, blood clotting and vasoconstriction) is a complex and redundant phenomenon, the salivary glands of blood sucking arthropods consist of a magic potion with diverse chemicals that in a redundant way counteract host mechanisms to prevent blood loss, allowing the fast acquisition of a meal. Salivary transcriptome made in the past few years indicate that the magic potion consists of 70-100 different proteins in the case of mosquitoes, for example, to over 1,000 in the case of ticks (Ticks feed for several days and have to disarm host immune reactions, in addition to the hemostatic system). Transcriptome studies also show that the salivary proteins of blood sucking arthropods are at a very fast pace of evolution, perhaps explaining why every genus studied so far has several unique protein families. Indeed, there are unique proteins found at the subgenus level. Given we can now describe in detail the sialotranscriptome (from the Greek word sialo = saliva) of a single organism, we can ask now what the universe of salivary proteins is associated to blood feeding, the so called sialoverse. There are near 19,000 species of blood sucking arthropods in 500 different genera. If we find (minimally) 5 novel protein families per genus (within the 70-500 proteins in each sialome), there are at least 2,500 novel proteins to be discovered, each one with an interesting pharmacological property. We have so far explored less than 20 genera of blood sucking arthropods, and it is our goal to extend sialotranscriptome discovery to map this pharmacological mine for future studies, and in the process learn the paths taken by genomes in their evolution to blood feeding and identify proteins with pharmacological and vaccine potential. While pursuing the theme above, Dr. Ribeiro has developed bioinformatic skills that were in high demand by laboratories within and outside NIAID, leading to many collaborative studies. During the 2019 fiscal year, members of the section of Vector Biology contributed to 12 publications, as described below: Transcriptome study: In the current fiscal year, we produced one paper related to sialotranscriptome of the tick Rhipicephalus sanguineus (1), a vector of rickettsiosis in the U.S.=. Transcriptomes and proteomes of the salivary glands of ticks fed to different degrees were analyzed, revealing the switching of proteins within the many protein families found in tick saliva. New insights into the mechanism of sialome switching were discussed. Functional study: We know for some years now that adult female mosquitoes have a unique salivary-expressed protein family, named the D7 family, which helps blood feeding by sequestering biogenic amines and leukotrienes. Surprisingly, we have found transcripts coding for a member of this protein family in fat body transcriptomes of both male and female mosquitoes. Dr. John Andersen took the lead on this project. Following recombinant expression of this protein, it was found that it binds juvenile hormone. Knock-out of the gene using crispr-cas9 methodology (done by Dr. Eric Calvo and Zachary Adelman, Virginia Tech) showed that this gene is important for the maturation of the innate immune response in adult Aedes aegypti mosquitoes (2). This study opens a new area of investigation related to the maturation of the immune response in mosquitoes. Bioinformatic collaborations: Dr. Ribeiro collaborated with intra and extra mural investigators lending his expertise in bioinformatics. With Drs. Patrick Duffy and Michal Fried (LMIV), using an assembly pipeline designed by Dr. Ribeiro, VAR protein coding transcripts from patient and culture-derived Plasmodium falciparum were successfully assembled and their predicted sequences verified by mass spectrometry (3-4). Novel VAR protein sequences (involved in the disease pathology and are vaccine targets) were discovered. This task of assembling transcripts of the highly repetitive VAR protein family was previously considered to be impossible. With a Brazilian group lead by Drs. Joao Santana Silva and Sandra Maruyama (State University of Sao Paulo, Brazil), we assembled and annotated the genome of protozoan kinetoplastids causing human disease similar to leishmaniasis, revealing a novel human parasite very similar to Crithidia fasciculata, a known parasite of flies that was not considered a parasite of mammals (5). We also collaborated with Dr. Saravanan Thangamani (SUNY Upstate Medical University) doing the assembly and annotation of tick salivary transcriptomes infected and non-infected with tick-borne encephalitis virus, at different times of feeding. Results revealed the virus-induced impairment of the salivary function, which delayed the sialome switching and possibly enhancing virus transmission (6). We additionally contributed to publications studying the transcriptional profiling and physiological roles of spermathecal-related genes in Aedes aegypti mosquitoes (in collaboration with Dr. Gustavo Martins, Universidade Federal de Vicosa, Brazil), (7) the transcription regulation in the embryonic development of mosquitoes and sand flies (8) (in collaboration with Dr. Urs Schmidt-Ott, University of Chicago), and the molecular mechanisms underlying milk production and viviparity in the cockroach, Diploptera punctata (9) where comparisons were made with the milk production in the ovoviviparous tsetse fly, a vector of African trypanosomiasis (in collaboration with Dr.Joshua Benoit, University of Cincinnati). Review articles Three review articles were produced during this fiscal year, reviewing the pharmacological properties of saliva from kissing bugs (10) (In collaboration with Dr. Jaime Santana, Federal University of Brasilia, Brazil), on the coevolution of vertebrates and blood sucking arthropods (11) (in collaboration with former LMVR fellows Anderson de S-Nunes, State University of So Paulo, Brazil and Carlos de Oliveira, Federal University of Triangulo Mineiro, Brazil), and on the salivary protein families of ticks, where a database was provided that will facilitate the task of tick salivary gland transcriptome annotation (12), which can produce over one thousand transcripts belonging to over 100 gene families.

由于宿主止血（防止失血的生理过程，包括血小板聚集、血液凝固和血管收缩）是一种复杂而多余的现象，吸血节肢动物的唾液腺由一种含有多种化学物质的神奇药水组成，这些化学物质以多余的方式抵消防止失血的宿主机制，允许快速获取食物。过去几年的唾液转录组表明，对于蚊子来说，这种神奇的药水由 70-100 种不同的蛋白质组成，例如，对于蜱虫来说，有超过 1,000 种蛋白质（蜱虫会进食数天，并且必须解除宿主的免疫反应），除了止血系统）。转录组研究还表明，吸血节肢动物的唾液蛋白进化速度非常快，这或许可以解释为什么迄今为止研究的每个属都有几个独特的蛋白质家族。事实上，在亚属水平上发现了独特的蛋白质。鉴于我们现在可以详细描述单个生物体的 sialotranscriptome（来自希腊语 sialo = 唾液），我们现在可以问唾液蛋白的宇宙与血液喂养有何关联，即所谓的 sialoverse。吸血节肢动物有 500 个不同属，近 19,000 种。如果我们（至少）在每个属中发现 5 个新蛋白质家族（每个 sialome 中的 70-500 个蛋白质），则至少有 2,500 个新蛋白质有待发现，每个蛋白质都具有有趣的药理学特性。到目前为止，我们已经探索了不到 20 个吸血节肢动物属，我们的目标是扩展唾液转录组的发现，为未来的研究绘制这个药理学矿藏，并在此过程中了解基因组在进化到吸血的过程中所采取的路径，并识别具有药理学和疫苗潜力的蛋白质。在追求上述主题的同时，Ribeiro 博士开发了 NIAID 内外实验室急需的生物信息技能，从而促成了许多合作研究。在 2019 财年，媒介生物学部门的成员发表了 12 篇出版物，如下所述：转录组研究：在本财年，我们发表了一篇与血红扇头蜱 (Rhipicephalus sanguineus) 唾液转录组相关的论文，该蜱是美国立克次体病的传播媒介=。对不同程度喂养的蜱唾液腺的转录组和蛋白质组进行了分析，揭示了蜱唾液中发现的许多蛋白质家族中蛋白质的转换。讨论了对唾液酸开关机制的新见解。功能研究：多年来，我们知道成年雌性蚊子有一个独特的唾液表达蛋白家族，称为 D7 家族，它通过隔离生物胺和白三烯来帮助血液喂养。令人惊讶的是，我们在雄性和雌性蚊子的脂肪体转录组中发现了编码该蛋白质家族成员的转录本。约翰·安德森博士领导了这个项目。该蛋白重组表达后，发现它与保幼激素结合。使用 Cripr-cas9 方法（由弗吉尼亚理工大学的 Eric Calvo 博士和 Zachary Adelman 完成）敲除该基因表明，该基因对于成年埃及伊蚊先天免疫反应的成熟非常重要 (2)。这项研究开辟了一个与蚊子免疫反应成熟相关的新研究领域。生物信息学合作：里贝罗博士与校内和校外的研究人员合作，发挥了他在生物信息学方面的专业知识。与博士。 Patrick Duffy 和 Michal Fried (LMIV) 使用 Ribeiro 博士设计的组装管道，成功组装了来自患者和培养物来源的恶性疟原虫的 VAR 蛋白编码转录物，并通过质谱验证了其预测序列 (3-4)。发现了新的 VAR 蛋白序列（涉及疾病病理学并且是疫苗靶标）。组装高度重复的 VAR 蛋白家族转录本的任务以前被认为是不可能的。与博士领导的巴西小组一起。 Joao Santana Silva 和 Sandra Maruyama（巴西圣保罗州立大学），我们组装并注释了导致类似于利什曼病的人类疾病的原生动物动质体的基因组，揭示了一种与束短线虫非常相似的新型人类寄生虫，束线虫是一种已知的苍蝇寄生虫，不被视为哺乳动物的寄生虫 (5)。我们还与 Saravanan Thangamani 博士（纽约州立大学上州医科大学）合作，在不同的喂养时间对感染和未感染蜱传脑炎病毒的蜱唾液转录组进行组装和注释。结果揭示了病毒引起的唾液功能损伤，从而延迟了唾液酸转换并可能增强病毒传播 (6)。我们还为研究埃及伊蚊受精囊相关基因的转录谱和生理作用的出版物做出了贡献（与巴西维科萨联邦大学的 Gustavo Martins 博士合作），(7) 蚊子胚胎发育中的转录调控和白蛉 (8)（与芝加哥大学 Urs Schmidt-Ott 博士合作），以及产奶和胎生的分子机制蟑螂，Diploptera punctata (9)，与非洲锥虫病媒介卵胎生采采蝇的产奶量进行了比较（与辛辛那提大学的 Joshua Benoit 博士合作）。评论文章本财年发表了三篇综述文章，综述了接吻虫唾液的药理学特性 (10)（与巴西巴西利亚联邦大学 Jaime Santana 博士合作），以及脊椎动物和吸血节肢动物的共同进化 (11) ）（与巴西圣保罗州立大学前 LMVR 研究员 Anderson de S-Nunes 和 Triangulo 联邦大学 Carlos de Oliveira 合作Mineiro，巴西），以及蜱的唾液蛋白家族，其中提供了一个数据库，该数据库将有助于蜱唾液腺转录组注释的任务（12），它可以产生属于 100 多个基因家族的 1000 多个转录本。